Abstracts

S17

ity (PWV), a non-invasive index of arterial stiffness and marker of cardiovascular risk in healthy
children.
Methods: In 573 healthy children (mean age 10.1 0.3
years; 51% boys), serum homocysteine level was measured by uorescent polarization immunoassay. PWV
was assessed noninvasively by applanation tonometry (Sphygmocor, AtCor Medical, Sydney, Australia).
Pubertal development was determined by Tanner stage.
Adiposity was assessed by percentage body fat (%BF)
using dual-energy X-ray absorptiometry. Insulin resistance was assessed by homeostasis model assessment (HOMA-IR) using fasting insulin and glucose
levels.
Results: Homocysteine was positively correlated with
Tanner stage (rho = 0.158, p = 0.043), %BF (rho = 0.146,
p = 0.020) and PWV (rho = 0.160, p = 0.009) in girls but not
boys. After adjustment for age, systolic blood pressure,
mean arterial pressure, heart rate, HOMA-IR, % BF and
Tanner stage; homocysteine was associated with PWV in
girls (p = 0.017).
Conclusion: Homocysteine was positively related to
arterial stiffness assessed by PWV, although the relation
was signicant only in girls. The signicance and mechanisms underlying the apparent sexual dimorphism in the
relationship between homocysteine and PWV require further evaluation.

status of the regulatory Thr855-MYPT subunit of myosin

phosphatase.
Results: In vessels with an intact endothelium simvastatin attenuated 40% of thromboxane A2 receptormediated constriction (P < 0.05) and increased phosphorylation of Ser1177-eNOS by 60% (P < 0.05). Interestingly,
simvastatin maintained 80% of its vasodilatory properties in the absence of an endothelium or in the
presence of a NOS inhibitor (L-NAME). In the
absence of endothelium, thromboxane-A2 receptor activation increased the inhibitory phosphorylation of myosin
phosphatase (Thr855-MYPT) by 30% (P < 0.05). Simvastatin or direct Rho kinase inhibition attenuated
the thromboxane-A2 receptor mediated phosphorylation
of MYPT.
Conclusions: Acute treatment with simvastatin attenuates thromboxane-A2 receptor-mediated vasoconstriction
via two mechanisms: (1) an endothelial/nitric oxidedependent mechanism and (2) increased activation of
myosin phosphatase.

doi:10.1016/j.hlc.2010.06.702

BakerIDI Heart and Diabetes Institute, Australia

36

The sympathetic nervous system (SNS) is central to the

regulation of blood pressure. SNS function can be divided
into 3 components: electrical (nerve ring), neurochemical (noradrenalin (NA) release) and cellular (regulatory
proteins in the sympathetic varicosity). Electrical and
biochemical SNS function can be measured utilising
microneurography and catecholamine kinetics; however,
there is no methodology for studying SNS function at a cellular level. We report on the development of sympathetic
nerve protein analysis in patients with recurrent vasovagal
syncope.
Results: We systematically assessed SNS function in
healthy subjects (n = 18) and patients with VVS (n = 45).
The VVS subjects fall into 2 groups, low supine systolic
BP (LSBP, n = 20) and normal BP (NSBP, n = 25). A subset of
patients underwent forearm vein biopsy for SNS protein
analysis. The proteins quantied were tyrosine hydroxylase (TH), the rate limiting enzyme in catecholamine
biosynthesis, norepinephrine transporter (NET), vesicular
monoamine transporter 2 and dynamin 1. LSBP subjects had paradoxically high nerve ring and low release
of NA; a nding attributed to reduced synthesis due
to markedly reduced TH expression. In contrast, NSBP
subjects had normal nerve ring rates and TH expression but reduced NA release attributable to high NET
expression.

Simvastatin Modulates Vasoconstriction via Activation

of Smooth Muscle Myosin Phosphatase and Endothelial
Nitric Oxide Synthase
S. Copley 1,2,3, , J. Beltrame 1,2,3 , D. Wilson 1,2,3
1 The

University of Adelaide, Australia

Elizabeth Hospital, Adelaide, Australia
3 The Basil Hetzel Institute, Adelaide, Australia
2 Queen

Introduction: Statins attenuate vasoconstriction independent from their lipid lowering properties. Using intact
vessels, in an acute setting, we examined two RhoA/Rho
kinase dependent mechanisms that may account for this
effect: (1) activation of endothelial nitric oxide synthase
(eNOS) and (2) activation of smooth muscle myosin phosphatase.
Methods: Rat caudal artery segments with/without
intact endothelium were mounted in a wire-myograph.
Arteries were incubated with simvastatin (60 nM to
10 mM) and/or the Rho kinase inhibitor H1152 (1 M)
and/or the nitric oxide synthase inhibitor, L-NAME
(10 M) for 60 min. Vessels were then constricted with
the thromboxane-A2 receptor agonist, U46619 (1 M)
and snap frozen to enable biochemical analysis. The
activation state of eNOS was assessed using ratiometric western blot analysis of the phosphorylation state
of Ser1177-eNOS. Rho kinase and myosin phosphatase
activity were similarly assessed for the phosphorylation

doi:10.1016/j.hlc.2010.06.703
37
Sympathetic Nerve Protein Analysis: A Novel Window
into Sympathetic Nervous System Dysfunction
G. Vaddadi , E. Lambert, L. Guo, T. Dawood, M. Esler

ABSTRACTS

Heart, Lung and Circulation

2010;19S:S1S268